Extraction of secondary metabolites for HPLC analysis
The biological material for the analysis
Inoculate three Bells plates with agar plugs (4 – 8 mm) and incubate for 14 days at 20 oC in darkness.
(1) Bell, A.A. et al. Pest Management Science 59, 6-7 (2003): 736-747.
(2) Smedsgaard J. Journal of Chromatography A 760.2 (1997): 264-270.
Filamentous fungi are notorious for their production of toxic metabolites (with both acute and cumulative effects). For most of these the effects of long time exposure have not been examined and it is therefore a good idea to use gloves (latex or nitrile) when working directly with the mycelia or used growth media.
Many of the chemicals used for this protocol is highly toxic (organic solvents) and great care should therefore be taken. Always use H4 gloves (or similar) when you are working with the “extraction solution” and “methanol”. All work should be performed in a fume hood to prevent inhalation of the chemicals. The use of gloves also protect against the fungal metabolites, which are very mobile in the used solvents, meaning that they will penetrate the skin easily.
Dette sted blev sidst opdateret 14. July 2010